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1.
ACS Synth Biol ; 2024 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-39292964

RESUMO

Phosphatidylinositol (PI) and its phosphorylated derivatives are of paramount importance in cellular functions and diseases. Understanding their diverse roles is, however, challenged by difficulties in synthesis and labeling techniques. In this proof-of-concept study, we demonstrate that PI can be straightforwardly de novo-synthesized and deuterium (2H)-labeled in Escherichia coli by genomic insertion of PI synthase from Trypanosoma brucei under constitutive synthetic promoter proD. Insertion into loci atpi-gidB and ybb revealed PI accumulation of 41% and 34% (mol/mol), respectively, when cultivated with glycerol as the sole carbon source. Growth of the atpi-gidB-PIS strain in deuterium-labeled (2H) substrates D2O, D8-glycerol, and D6-myo-inositol achieved PI deuteration of 90%, PE deuteration of 95%, and total fatty acids|fatty acid (FA) deuteration of 97%. This study offers an alternative convenient route to chemical and enzymatic labeling synthesis of PI; more excitingly, this work also, in principle, opens a door for tailoring the FA profile of deuterated PI/PE for task-specific application by repurposing FA biosynthesis pathways.

2.
Microsc Res Tech ; 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-39237471

RESUMO

The development of new treatments for ocular diseases often requires investigating eyes similar in size and structure to human eyes. Such studies are challenging because analyzing the histopathology of large, human-sized eyes can be technically difficult. In particular, obtaining high-quality frozen sections is almost impossible due to the formation of ice crystals in the vitreous, which causes crush artifacts during the procedures of section and post sectioning manipulations. Herein, we describe a new method that provides high-quality frozen sections for large eyes and demonstrate its usefulness in the eyes of rabbits, pigs, minipigs, monkeys, and humans. We observed that artifactual separation of the photoreceptors from the retinal pigment epithelium is minimized and photoreceptor morphology is preserved. This method can be highly beneficial for investigators seeking to translate new treatments for ocular disease into the clinic. RESEARCH HIGHLIGHTS: Histopathological analysis of large and human-sized eyes presents significant challenges, particularly in obtaining high-quality frozen sections. A multistep fixation followed by vitreous removal and replacement ensures better cryopreservation and embedding of large eyes, minimizing the morphological and structural retinal loss found in many studies. Our results demonstrate that a multistep fixation and cryopreservation method for large eyes in histopathology consistently minimizes the artifactual separation of photoreceptors from the retinal pigment epithelium, thereby preserving photoreceptor morphology and providing high-quality frozen sections. A new method providing high-quality sections is necessary and will be highly useful for investigators aiming to translate new treatments for ocular diseases into clinical applications.

3.
Int J Biol Macromol ; 278(Pt 2): 134640, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39142484

RESUMO

The successful germination of pollen is essential for double fertilization in flowering plants. Mechanosensitive channels of small conductance (MscS-like, MSL) inhibit pollen germination and maintains cellular integrity of pollen during this process. Therefore, it is vital to carefully regulate the expression of MSL to promote successful pollen germination. Despite its importance, the molecular mechanisms governing MSL expression in plants remain poorly understood. Here, we had identified 15 MSL genes in the pear, among which PbrMSL5 was expressed in pollen development. Subcellular localization experiments revealed that PbrMSL5 was located in both plasma membrane and cytoplasm. Functional investigations, including complementation experiments using the atmsl8 mutant background, demonstrated the involvement of PbrMSL5 in preserving pollen cell integrity and inhibiting germination. Antisense oligonucleotide experiments further confirmed that PbrMSL5 suppressed pear pollen germination by reducing osmotic pressure and Cl- content. Yeast one-hybrid, electrophoretic mobility shift assays, and dual luciferase reporter assay elucidated that PbrMYC8 interacts directly with the N-box element, leading to the suppression of PbrMSL5 expression and promoted pollen germination. These results represented a significant advancement in unraveling the molecular mechanisms controlling plant MSL expression. This study showed valuable contribution to advancing our comprehension of the mechanism underlying pollen germination.


Assuntos
Regulação da Expressão Gênica de Plantas , Germinação , Proteínas de Plantas , Pólen , Pyrus , Fatores de Transcrição , Pólen/genética , Germinação/genética , Pyrus/genética , Pyrus/metabolismo , Pyrus/crescimento & desenvolvimento , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
4.
J Colloid Interface Sci ; 678(Pt A): 593-601, 2024 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-39216387

RESUMO

Paper-based relics is an important carrier for recording and preserving information, however, it faces irreversible UV-induced damage, including photocleavage, oxidation, acidification and discoloration, which seriously affects its value and lifespan. Carbon dots (CDs) possess excellent UV absorption and good chemical stability, making them suitable for UV protection. Herein, we propose a high-security and efficient method utilizing CDs films (CDFs) for preventive protection of paper against UV damage. The CDFs with high tunable UV absorbance and minimal absorbance in the visible light range, effectively shield paper from UV radiation while preserving its visual appeal. Moreover, the UV transmittance of the film can be fine-tuned to the content of CDs and can be easily removed from the paper without residue. Artificial accelerated UV aging experiments demonstrate the deceleration of acidification, oxidation, and photocleavage in the protected bamboo paper and Xuan paper. This research paces a new direction for the protection of paper and paper-based relics and artworks with emerging carbon materials, offering customizable protection effects tailored to specific preservation and exhibition requirements. This research pioneers a novel approach to preventive protection of paper and paper-based relics using emerging carbon dots materials, offering tailored protection for diverse preservation needs.

5.
Entropy (Basel) ; 26(8)2024 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-39202103

RESUMO

Random matrix series are a significant component of random matrix theory, offering rich theoretical content and broad application prospects. In this paper, we propose modified versions of tail bounds for random matrix series, including matrix Gaussian (or Rademacher) and sub-Gaussian and infinitely divisible (i.d.) series. Unlike present studies, our results depend on the intrinsic dimension instead of ambient dimension. In some cases, the intrinsic dimension is much smaller than ambient dimension, which makes the modified versions suitable for high-dimensional or infinite-dimensional setting possible. In addition, we obtain the expectation bounds for random matrix series based on the intrinsic dimension.

6.
J Hazard Mater ; 478: 135493, 2024 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-39173381

RESUMO

Aflatoxin is one of the most notorious mycotoxins, of which aflatoxin B1 (AFB1) is the most harmful and prevalent. Microbes play a crucial role in the environment for the biotransformation of AFB1. In this study, a bacterial consortium, HS-1, capable of degrading and detoxifying AFB1 was obtained. Here, we combined multi-omics and cultivation-based techniques to elucidate AFB1 biotransformation by consortium HS-1. Co-occurrence network analysis revealed that the key taxa responsible for AFB1 biotransformation in consortium HS-1 mainly belonged to the phyla Proteobacteria and Actinobacteria. Moreover, metagenomic analysis showed that diverse microorganisms, mainly belonging to the phyla Proteobacteria and Actinobacteria, carry key functional enzymes involved in the initial step of AFB1 biotransformation. Metatranscriptomic analysis indicated that Paracoccus-related bacteria were the most active in consortium HS-1. A novel bacterium, Paracoccus sp. strain XF-30, isolated from consortium HS-1, contains a novel dye-decolorization peroxidase (DyP) enzyme capable of effectively degrading AFB1. Taxonomic profiling by bioinformatics revealed that DyP, which is involved in the initial biotransformation of AFB1, is widely distributed in metagenomes from various environments, primarily taxonomically affiliated with Proteobacteria and Actinobacteria. The in-depth examination of AFB1 biotransformation in consortium HS-1 will help us to explore these crucial bioresources more sensibly and efficiently.


Assuntos
Actinobacteria , Aflatoxina B1 , Biotransformação , Proteobactérias , Aflatoxina B1/metabolismo , Actinobacteria/metabolismo , Actinobacteria/genética , Proteobactérias/metabolismo , Proteobactérias/genética , Paracoccus/metabolismo , Paracoccus/genética , Biodegradação Ambiental
7.
J Ethnopharmacol ; 335: 118600, 2024 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-39053714

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Herb-induced liver injury (HILI) represents an exacerbated inflammatory response, with Psoraleae fructus (PF) and its preparations recently associated with hepatotoxicity. Licorice, historically recognized as a detoxifying herbal remedy, is considered to possess hepatoprotective properties. Our previous research identified bavachin, bakuchiol, and psoralidin (PSO) as potential toxic constituents in PF, while licochalcone B (LCB) and echinatin were identified as bioactive components in licorice. However, evidence regarding the interactions of active compounds in herbs and their underlying mechanisms remains limited. AIM OF THE STUDY: The objective of this study is to assess the potential mechanisms through which LCB modulates immunological and anti-inflammatory responses to treat PSO-induced liver injury by using human hepatocyte cells (L02) and LPS-primed mice. METHODS: The ameliorative effects of LCB and echinatin on bavachin, bakuchiol, and PSO-induced liver injury were demonstrated in L02 cells. Subsequently, the efficacy of LCB on PSO-induced idiosyncratic liver injury was further validated in C57BL/6 mice under moderate inflammatory stress induced by LPS priming. The mechanisms were preliminarily explored with an integrated strategy of molecular docking, RT-PCR verification, and untargeted metabolomics. RESULTS: The study shows that LCB significantly reduced cell injury induced by the three chemicals in PF and provided substantial protection against PSO-induced hepatic damage, as indicated by the levels of ALT, AST, and LDH. LCB normalized liver function and remarkedly alleviated hepatic lesions and inflammation caused by PSO in mice under moderate inflammatory stress. The mRNA profiles of both L02 cells and mice liver tissue revealed that LCB mitigated PSO-induced hepatotoxicity by regulating the gene expression of pro-inflammatory cytokines IL1B and TNF, as well as immunoinflammatory genes PIK3CA, AKT1, NFKB1, and NLRP3. Furthermore, untargeted metabolomics of liver tissue indicated that LCB could reverse the abnormal expression of 11 discriminatory metabolites, with the interrelationship between differential metabolites and target genes primarily clustering in glycerophospholipid metabolism, arachidonic acid metabolism, and phosphatidylinositol signaling system. CONCLUSION: LCB demonstrated a superior anti-inflammatory and immunomodulatory effect on PSO-induced hepatotoxicity by modulating the inflammatory response and metabolic signaling system. Key interactive targets included phosphatidylcholine, phosphatidic acid, and subunit isoforms of PI3K.


Assuntos
Anti-Inflamatórios , Benzofuranos , Chalconas , Doença Hepática Induzida por Substâncias e Drogas , Camundongos Endogâmicos C57BL , Fenóis , Animais , Chalconas/farmacologia , Anti-Inflamatórios/farmacologia , Humanos , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Doença Hepática Induzida por Substâncias e Drogas/patologia , Masculino , Benzofuranos/farmacologia , Fenóis/farmacologia , Camundongos , Linhagem Celular , Cumarínicos/farmacologia , Fatores Imunológicos/farmacologia , Simulação de Acoplamento Molecular , Glycyrrhiza/química , Lipopolissacarídeos/toxicidade , Fígado/efeitos dos fármacos , Fígado/patologia , Fígado/metabolismo , Hepatócitos/efeitos dos fármacos , Citocinas/metabolismo , Flavonoides/farmacologia
8.
Chemosphere ; 362: 142564, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38885762

RESUMO

Atmospheric pollution has been demonstrated to be associated with ocular surface diseases characterized by corneal epithelial damage, including impaired barrier function and squamous metaplasia. However, the specific mechanisms underlying the impact of atmospheric pollution on corneal damage are still unknow. To address this gap in knowledge, we conducted a study using a whole-body exposure system to investigate the detrimental effects of traffic-related air pollution, specifically diesel exhaust (DE), on corneal epithelium in C57BL/6 mice over a 28-day period. Following DE exposure, the pathological alterations in corneal epithelium, including significant increase in corneal thickness and epithelial stratification, were observed in mice. Additionally, exposure to DE was also shown to disrupt the barrier functions of corneal epithelium, leading to excessive proliferation of basal cells and even causing squamous metaplasia in corneal epithelium. Further studies have found that the activation of yes-associated protein (YAP), characterized by nuclear translocation, may play a significant role in DE-induced corneal squamous metaplasia. In vitro assays confirmed that DE exposure triggered the YAP/ß-catenin pathway, resulting in squamous metaplasia and destruction of barrier functions. These findings provide the preliminary evidence that YAP activation is one of the mechanisms of the damage to corneal epithelium caused by traffic-related air pollution. These findings contribute to the knowledge base for promoting eye health in the context of atmospheric pollution.


Assuntos
Poluentes Atmosféricos , Epitélio Corneano , Metaplasia , Camundongos Endogâmicos C57BL , Emissões de Veículos , Proteínas de Sinalização YAP , Emissões de Veículos/toxicidade , Animais , Epitélio Corneano/efeitos dos fármacos , Epitélio Corneano/patologia , Camundongos , Poluentes Atmosféricos/toxicidade , Masculino , beta Catenina/metabolismo , Proliferação de Células/efeitos dos fármacos
9.
J Agric Food Chem ; 72(23): 13297-13307, 2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38830127

RESUMO

2-(2-Phenylethyl)chromones (PECs) are the primary constituents responsible for the promising pharmacological activities and unique fragrance of agarwood. However, the O-methyltransferases (OMTs) involved in the formation of diverse methylated PECs have not been reported. In this study, we identified one Mg2+-dependent caffeoyl-CoA-OMT subfamily enzyme (AsOMT1) and three caffeic acid-OMT subfamily enzymes (AsOMT2-4) from NaCl-treated Aquilaria sinensis calli. AsOMT1 not only converts caffeoyl-CoA to feruloyl-CoA but also performs nonregioselective methylation at either the 6-OH or 7-OH position of 6,7-dihydroxy-PEC. On the other hand, AsOMT2-4 preferentially utilizes PECs as substrates to produce structurally diverse methylated PECs. Additionally, AsOMT2-4 also accepts nonPEC-type substrates such as caffeic acid and apigenin to generate methylated products. Protein structure prediction and site-directed mutagenesis revealed that residues of L313 and I318 in AsOMT3, as well as S292 and F313 in AsOMT4 determine the distinct regioselectivity of these two OMTs toward apigenin. These findings provide important biochemical evidence of the remarkable structural diversity of PECs in agarwood.


Assuntos
Metiltransferases , Proteínas de Plantas , Thymelaeaceae , Metiltransferases/genética , Metiltransferases/química , Metiltransferases/metabolismo , Thymelaeaceae/enzimologia , Thymelaeaceae/química , Thymelaeaceae/genética , Proteínas de Plantas/genética , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Madeira/química , Especificidade por Substrato , Ácidos Cafeicos/química , Ácidos Cafeicos/metabolismo , Metilação , Flavonoides
10.
Mol Cell Biochem ; 2024 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-38880861

RESUMO

Acute myocardial infarction is mainly caused by a lack of blood flood in the coronary artery. Angiopoietin-like protein 2 (ANGPTL2) induces platelet activation and thrombus formation in vitro through binding with immunoglobulin-like receptor B, an immunoglobulin superfamily receptor. However, the mechanism by which it regulates platelet function in vivo remains unclear. In this study, we investigated the role of ANGPTL2 during thrombosis in relationship with ST-segment elevation myocardial infarction (STEMI) with spontaneous recanalization (SR). In a cohort of 276 male and female patients, we measured plasma ANGPTL2 protein levels. Using male Angptl2-knockout and wild-type mice, we examined the inhibitory effect of Angptl2 on thrombosis and platelet activation both in vivo and ex vivo. We found that plasma and platelet ANGPTL2 levels were elevated in patients with STEMI with SR compared to those in non-SR (NSR) patients, and was an independent predictor of SR. Angptl2 deficiency accelerated mesenteric artery thrombosis induced by FeCl3 in Angptl2-/- compared to WT animals, promoted platelet granule secretion and aggregation induced by thrombin and collogen while purified ANGPTL2 protein supplementation reversed collagen-induced platelet aggregation. Angptl2 deficiency also increased platelet spreading on immobilized fibrinogen and clot contraction. In collagen-stimulated Angptl2-/- platelets, Src homology region 2 domain-containing phosphatase (Shp)1-Y564 and Shp2-Y580 phosphorylation were attenuated while Src, Syk, and Phospholipase Cγ2 (PLCγ2) phosphorylation increased. Our results demonstrate that ANGPTL2 negatively regulated thrombus formation by activating ITIM which can suppress ITAM signaling pathway. This new knowledge provides a new perspective for designing future antiplatelet aggregation therapies.

11.
J Hazard Mater ; 476: 135045, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-38944990

RESUMO

Isoprocarb (IPC), a representative monocyclic carbamate insecticide, poses risks of environmental contamination and harm to non-target organisms. However, its degradation mechanism has not been reported. In this study, a newly IPC-degrading strain D-6 was isolated from the genus Rhodococcus, and its degradation characteristics and pathway of IPC were analyzed. A novel hydrolase IpcH, responsible for hydrolyzing IPC to 2-isopropylphenol (IPP), was identified. IpcH exhibited low similarity (< 27 %) with other reported hydrolases, including previously characterized carbamate insecticides hydrolases, indicating its novelty. The Km and kcat values of IpcH towards IPC were 69.99 ± 8.33 µM and 95.96 ± 4.02 s-1, respectively. Also, IpcH exhibited catalytic activity towards various types of carbamate insecticides, including monocyclic carbamates (IPC, fenobucarb and propoxur), bicyclic carbamates (carbaryl and carbofuran), and linear carbamates (oxamyl and aldicarb). The molecular docking and site-directed mutagenesis revealed that His254, His256, His329 and His376 were essential for IpcH activity. Strain D-6 can effectively reduce the toxicity of IPC and IPP towards sensitive organisms through its degradation ability. This study presents the initial report on IPC degradation pathway and molecular mechanism of IPC degradation, and provides a good potential strain for bioremediating IPC and IPP-contaminated environments.


Assuntos
Biodegradação Ambiental , Hidrolases , Inseticidas , Rhodococcus , Rhodococcus/metabolismo , Rhodococcus/genética , Hidrolases/metabolismo , Hidrolases/genética , Inseticidas/metabolismo , Inseticidas/química , Inseticidas/toxicidade , Simulação de Acoplamento Molecular , Uretana/metabolismo , Uretana/química
12.
J Hazard Mater ; 476: 134968, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-38901263

RESUMO

The widely used phenylurea herbicide isoproturon (IPU) and its residues can inhibit the growth of subsequently planted crops. However, reports on bioremediation of IPU-contaminated soil are scarce. In this study, Sphingobium sp. strain YBL2-gfp (a derivative of the IPU-degrading Sphingobium sp. strain YBL2 isolated by our lab) was constructed to bioremediate IPU-contaminated soil. In pot experiments, strain YBL2-gfp colonized the roots of wheat and eliminated IPU residues in the soil within 21 d, effectively alleviating its toxicity and restoring wheat growth. IPU treatment reduced the richness and diversity of soil bacteria, while inoculation YBL2-gfp mainly affected richness with less impact on diversity. The high concentrations of IPU and inoculation of YBL2-gfp alone reduced the soil microbial community connections, while bioaugmentation treatment enhanced the soil microbial community connections. Additionally, strain YBL2-gfp stimulated the metabolic capacity of the indigenous microbes, promoting the degradation of IPU and reducing the negative impact of high concentrations of IPU on microbial community. Taken together, this study offers relatively comprehensive insights into the practical application of bioaugmentation, demonstrating that strain YBL2 has the potential to remediate IPU-contaminated soils.


Assuntos
Biodegradação Ambiental , Herbicidas , Compostos de Fenilureia , Microbiologia do Solo , Poluentes do Solo , Sphingomonadaceae , Poluentes do Solo/metabolismo , Sphingomonadaceae/metabolismo , Sphingomonadaceae/genética , Sphingomonadaceae/crescimento & desenvolvimento , Herbicidas/metabolismo , Compostos de Fenilureia/metabolismo , Triticum/metabolismo , Triticum/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento
13.
Cell Biochem Funct ; 42(4): e4020, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38702967

RESUMO

The regulatory potential of long noncoding RNA (lncRNA) FBXL19-AS1 has been highlighted in various cancers, but its effect on triple-negative breast cancer (TNBC) remains unclear. Here, we aimed to elucidate the role of FBXL19-AS1 in TNBC and its underlying mechanism. RT-qPCR was employed to detect the expressions of FBXL19-AS1 and miR-378a-3p in tissues and cells. Immunohistochemical staining and western blot were utilized to detect the expression levels of proteins. Cell activities were detected using flow cytometry, CCK-8, and transwell assay. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were deployed to investigate interactions of different molecules. Protein-protein interaction (PPI) network, gene ontology (GO), and Kyoto encyclopedia of genes and genomes (KEGG) pathways were used to analyze the downstream pathway. In vivo xenograft model was conducted to detect the effect of FBXL19-AS1 on tumor growth. FBXL19-AS1 was overexpressed in TNBC tissues and cell lines compared with counterparts. FBXL19-AS1 knockdown suppressed TNBC cell activities, whereas its overexpression exhibited the opposite effect. Mechanistically, FBXL19-AS1 was found to interact with miR-378a-3p. Further analysis revealed that miR-378a-3p exerted tumor-suppressive effects in TNBC cells. Additionally, miR-378a-3p targeted and downregulated the expression of ubiquitin aldehyde binding 2 (OTUB2), a deubiquitinase associated with TNBC progression. In vivo experiments substantiated the inhibitory effects of FBXL19-AS1 knockdown on TNBC tumorigenesis, and a miR-378a-3p inhibitor partially rescued these effects. The downstream pathway of the miR-378a-3p/OTUB2 axis was explored, revealing connections with proteins involved in modifying other proteins, removing ubiquitin molecules, and influencing signaling pathways, including the Hippo signaling pathway. Western blot analysis confirmed changes in YAP and TAZ expression levels, indicating a potential regulatory network. In summary, FBXL19-AS1 promotes exacerbation in TNBC by suppressing miR-378a-3p, leading to increased OTUB2 expression. The downstream mechanism may be related to the Hippo signaling pathway. These findings propose potential therapeutic targets for TNBC treatment.


Assuntos
MicroRNAs , RNA Longo não Codificante , Neoplasias de Mama Triplo Negativas , Animais , Feminino , Humanos , Camundongos , Linhagem Celular Tumoral , Proliferação de Células , Enzimas Desubiquitinantes/metabolismo , Proteínas F-Box/metabolismo , Proteínas F-Box/genética , Regulação Neoplásica da Expressão Gênica , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/metabolismo , MicroRNAs/genética , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologia , Neoplasias de Mama Triplo Negativas/genética
14.
Appl Environ Microbiol ; 90(5): e0197423, 2024 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-38619269

RESUMO

17ß-estradiol (E2) is a natural endocrine disruptor that is frequently detected in surface and groundwater sources, thereby threatening ecosystems and human health. The newly isolated E2-degrading strain Sphingomonas colocasiae C3-2 can degrade E2 through both the 4,5-seco pathway and the 9,10-seco pathway; the former is the primary pathway supporting the growth of this strain and the latter is a branching pathway. The novel gene cluster ean was found to be responsible for E2 degradation through the 4,5-seco pathway, where E2 is converted to estrone (E1) by EanA, which belongs to the short-chain dehydrogenases/reductases (SDR) superfamily. A three-component oxygenase system (including the P450 monooxygenase EanB1, the small iron-sulfur protein ferredoxin EanB2, and the ferredoxin reductase EanB3) was responsible for hydroxylating E1 to 4-hydroxyestrone (4-OH-E1). The enzymatic assay showed that the proportion of the three components is critical for its function. The dioxygenase EanC catalyzes ring A cleavage of 4-OH-E1, and the oxidoreductase EanD is responsible for the decarboxylation of the ring A-cleavage product of 4-OH-E1. EanR, a TetR family transcriptional regulator, acts as a transcriptional repressor of the ean cluster. The ean cluster was also found in other reported E2-degrading sphingomonads. In addition, the novel two-component monooxygenase EanE1E2 can open ring B of 4-OH-E1 via the 9,10-seco pathway, but its encoding genes are not located within the ean cluster. These results refine research on genes involved in E2 degradation and enrich the understanding of the cleavages of ring A and ring B of E2.IMPORTANCESteroid estrogens have been detected in diverse environments, ranging from oceans and rivers to soils and groundwater, posing serious risks to both human health and ecological safety. The United States National Toxicology Program and the World Health Organization have both classified estrogens as Group 1 carcinogens. Several model organisms (proteobacteria) have established the 4,5-seco pathway for estrogen degradation. In this study, the newly isolated Sphingomonas colocasiae C3-2 could degrade E2 through both the 4,5-seco pathway and the 9,10-seco pathway. The novel gene cluster ean (including eanA, eanB1, eanC, and eanD) responsible for E2 degradation by the 4,5-seco pathway was identified; the novel two-component monooxygenase EanE1E2 can open ring B of 4-OH-E1 through the 9,10-seco pathway. The TetR family transcriptional regulator EanR acts as a transcriptional repressor of the ean cluster. The cluster ean was also found to be present in other reported E2-degrading sphingomonads, indicating the ubiquity of the E2 metabolism in the environment.


Assuntos
Biodegradação Ambiental , Estradiol , Família Multigênica , Sphingomonas , Sphingomonas/metabolismo , Sphingomonas/genética , Estradiol/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Disruptores Endócrinos/metabolismo , Filogenia
15.
Zygote ; 32(2): 175-182, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38629180

RESUMO

Intracytoplasmic sperm injection (ICSI) is a technique that directly injects a single sperm into the cytoplasm of mature oocytes. Here, we explored the safety of single-sperm cryopreservation applied in ICSI. This retrospective study enrolled 186 couples undergoing ICSI-assisted pregnancy. Subjects were allocated to the fresh sperm (group A)/single-sperm cryopreservation (group B) groups based on sperm type, with their clinical baseline/pathological data documented. We used ICSI-compliant sperm for subsequent in vitro fertilization and followed up on all subjects. The recovery rate/cryosurvival rate/sperm motility of both groups, the pregnancy/outcome of women receiving embryo transfer, and the delivery mode/neonatal-related information of women with successful deliveries were recorded. The clinical pregnancy rate, cumulative clinical pregnancy rate, abortion rate, ectopic pregnancy rate, premature delivery rate, live birth delivery rate, neonatal birth defect rate, and average birth weight were analyzed. The two groups showed no significant differences in age, body mass index, ovulation induction regimen, sex hormone [anti-Müllerian hormone (AMH)/follicle-stimulating hormone (FSH)/luteinizing hormone (LH)] levels, or oocyte retrieval cycles. The sperm recovery rate (51.72%-100.00%) and resuscitation rate (62.09% ± 16.67%) in group B were higher; the sperm motility in the two groups demonstrated no significant difference and met the ICSI requirements. Group B exhibited an increased fertilization rate, decreased abortion rate, and increased safety versus group A. Compared with fresh sperm, the application of single-sperm cryopreservation in ICSI sensibly improved the fertilization rate and reduced the abortion rate, showing higher safety.


Assuntos
Criopreservação , Taxa de Gravidez , Injeções de Esperma Intracitoplásmicas , Motilidade dos Espermatozoides , Espermatozoides , Humanos , Injeções de Esperma Intracitoplásmicas/métodos , Feminino , Criopreservação/métodos , Masculino , Gravidez , Adulto , Estudos Retrospectivos , Espermatozoides/fisiologia , Preservação do Sêmen/métodos , Resultado da Gravidez , Transferência Embrionária/métodos , Fertilização in vitro/métodos
16.
Bioorg Chem ; 147: 107362, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38615474

RESUMO

Excessive peroxynitrite (ONOO-) is closely related to the occurrence and progression of inflammation. Therefore, the development of an efficacious ONOO- activatable probe holds great potential for the early diagnosis of pathological inflammation, and the direct evaluation of the therapeutic efficacy of active protectants. In this work, a new ONOO--activated fluorescent probe (SZP) which greatly improved the specificity and sensitivity (LOD = 8.03 nM) with large Stokes shift (150 nm) through introducing two reaction triggers (diphenyl phosphinate moiety, CC unsaturated bond) was rationally designed for rapid detecting ONOO- (within 2 min). The excellent properties of probe SZP enable it to realize the fluorescence-guided diagnosis of inflammation. More importantly, probe SZP has also been utilized to assess the anti-inflammatory efficacy of traditional Chinese medicines (TCMs) active ingredients for the remediation of inflammation by monitoring ONOO- fluctuation for the first time.


Assuntos
Corantes Fluorescentes , Inflamação , Ácido Peroxinitroso , Ácido Peroxinitroso/análise , Ácido Peroxinitroso/antagonistas & inibidores , Corantes Fluorescentes/química , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/farmacologia , Inflamação/tratamento farmacológico , Animais , Estrutura Molecular , Camundongos , Humanos , Células RAW 264.7 , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/síntese química , Anti-Inflamatórios/uso terapêutico , Imagem Óptica , Relação Dose-Resposta a Droga , Relação Estrutura-Atividade , Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/síntese química , Masculino
17.
Asian J Surg ; 47(6): 2606-2612, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38548542

RESUMO

BACKGROUND: The accurate evaluation of surgical margins holds crucial importance in determining the success of breast-conserving surgery (BCS). The aim of this study was to introduce a novel technique for the positioning of surgical margins in BCS while highlighting its advantages. METHODS: This study included a cohort of breast cancer patients who underwent BCS. The patients were categorized into two groups: one group underwent BCS with the traditional palpation-guided method, and the other with the 3D-MPT technique. The study assessed and compared the feasibility, advantages, and outcomes in terms of quality of life between the two groups. RESULTS: A total of 80 patients were successfully enrolled in the study. No significant differences in clinicopathological features were observed between the two groups. The 3D-MPT technique was found to be feasible and offered several advantages over the palpation-guided method. The utilization of guide wires by experienced radiologists to position the margins before surgery enabled precise and swift specimen removal, resulting in the conservation of valuable time and a reduction in the need for re-excision. Furthermore, the 3D-MPT technique exhibited the potential to enhance cosmetic outcomes and elevate patient satisfaction, particularly in cases with uncertain tumor boundaries detectable by palpation. CONCLUSION: The 3D-MPT technique proves to be an effective and safe approach for reducing tumor positivity rates in initial surgical margins, thereby improving the quality of life for patients undergoing breast-conserving surgery in comparison to the conventional method.


Assuntos
Neoplasias da Mama , Estudos de Viabilidade , Margens de Excisão , Mastectomia Segmentar , Palpação , Qualidade de Vida , Humanos , Mastectomia Segmentar/métodos , Neoplasias da Mama/cirurgia , Neoplasias da Mama/patologia , Feminino , Palpação/métodos , Pessoa de Meia-Idade , Idoso , Adulto , Estudos de Coortes , Resultado do Tratamento , Imageamento Tridimensional , Satisfação do Paciente
18.
Sci Adv ; 10(10): eadl3576, 2024 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-38457512

RESUMO

Suprachoroidal nonviral gene therapy with biodegradable poly(ß-amino ester) nanoparticles (NPs) provides widespread expression in photoreceptors and retinal pigmented epithelial (RPE) cells and therapeutic benefits in rodents. Here, we show in a human-sized minipig eye that suprachoroidal injection of 50 µl of NPs containing 19.2 µg of GFP expression plasmid caused GFP expression in photoreceptors and RPE throughout the entire eye with no toxicity. Two weeks after injection of 50, 100, or 200 µl, there was considerable within-eye and between-eye variability in expression that was reduced 3 months after injection of 200 µl and markedly reduced after three suprachoroidal injections at different locations around the eye. Reduction of bacterial CpG sequences in the expression plasmid resulted in a trend toward higher expression. These data indicate that nonviral suprachoroidal gene therapy with optimized polymer, expression plasmid, and injection approach has potential for treating photoreceptors throughout the entire retina of a human-sized eye.


Assuntos
Nanopartículas , Retina , Animais , Humanos , Suínos , Porco Miniatura , Retina/metabolismo , Plasmídeos/genética , Terapia Genética/métodos
19.
Chemistry ; 30(31): e202400329, 2024 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-38551107

RESUMO

Green hydrogen production through electrochemical overall water splitting has suffered from sluggish oxygen evolution reaction (OER) kinetics, inferior conversion efficiency, and high cost. Herein, ultrafine PtIr clusters are synthesized via an electrodeposition method and decorated on the Co3O4 nanoflowers assembled by nanowires (PtIr-Co3O4). The encouraging performances in electrochemical OER and hydrogen evolution reaction (HER) are achieved over the PtIr-Co3O4 catalyst, with the overpotentials as low as 410 and 237 mV at 100 mA cm-2, respectively, outperforming the commercial IrO2 and Pt/C catalysts. Due to the ultralow loading of PtIr clusters, the PtIr-Co3O4 catalyst exhibits 1270 A gIr -1 for OER at the overpotential of 400 mV. Our detailed analyses also show that the strong interactions between the ultrafine PtIr clusters and the Co3O4 nanoflowers enable the PtIr-Co3O4 catalyst to afford 10 mA cm-2 for the overall water splitting at the potential of 1.57 V, accompanied by high durability for 100 h.

20.
J Chromatogr A ; 1720: 464782, 2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38442498

RESUMO

Semicarbazide, as a derivative of urea, constitutes a great variety of functional molecules for different needs. Herein, novel stationary phases with an incorporated semicarbazide group were proposed. Using aliphatic (docosanoyl, C22) and aromatic (benzoyl, Bz) hydrazides, the semicarbazide-embedded ligands were synthesized before chemical modification of the silica gel, allowing for an accurate interpretation of the chromatographic properties of the corresponding packings. The new stationary phases were water-wettable, due to the presence of highly polar groups. In particular, Bz-semicarbazide (Bz-SCD) stationary phase was sufficiently hydrophilic to run in hydrophilic interaction (HILIC) mode, whilst the C22 (C22-SCD) equivalent, in spite of its reversed-phase nature, was markedly less hydrophobic than the referenced polar-embedded ones. The versatility of C22-SCD was demonstrated with a large selection of analytes, including geometric isomers and standard mixtures of polycyclic aromatic hydrocarbons, sulfonamides, sulfonylurea, substituted ureas, pyridines and carbamates, fat-soluble colorants, antifungal metabolites, angiotensin II receptor blockers and calcium channel blockers.


Assuntos
Cromatografia de Fase Reversa , Semicarbazidas , Dióxido de Silício , Cromatografia Líquida/métodos , Interações Hidrofóbicas e Hidrofílicas , Isomerismo , Dióxido de Silício/química
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